Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Elevated PYGM (glycogen phosphorylase, muscle associated) expression in the astrocytes of tau P301S transgenic (PS19) mice at pathological stages and the brains of frontotemporal lobar degeneration with ubiquitin‐positive inclusions (FTLD‐U) patients. (A, B) Immunoblot (A) and quantitative analysis (B) of PYGM, PYGB, PYGL, and GYS1 protein levels in the hippocampus and cortex of 9‐month‐old male PS19 mice and wild‐type (WT) littermates. n = 8 per group. (C, D) Immunoblot (C) and quantitative analysis (D) of hippocampal PYGM protein levels in 2‐, 6‐, and 9‐month‐old male PS19 and WT littermates. n = 4 per group. (E, F) Immunofluorescence staining of PYGM (green) and the astrocytic marker glial fibrillary acidic protein (GFAP) (red) in the hippocampus and cortex of 9‐month‐old PS19 mice and WT littermates (E). PYGM‐positive area and PYGM colocalization with GFAP were quantified for comparison (F). The nuclei were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). Scale bars: 100 µm for regular and 20 µm for zoom‐in images. n = 8 per group. (G) Line scan analysis of PYGM and GFAP colocalization. The graph shows fluorescence intensity profiles of PYGM (blue) and GFAP (gray) along the distance of the rectangular region in the inset, indicating their spatial relationship. Scale bar: 20 µm. (H) Comparison of PYGM mRNA levels in post‐mortem brain tissues of FTLD‐U patients with progranulin gene mutations (progranulin‐mutant FTLD‐U, n = 15), FTLD‐U patients without progranulin gene mutations (sporadic FTLD‐U, n = 24), and healthy controls (control, n = 17) from the GSE13162 dataset. Data are presented as mean ± SEM. p values were determined by two‐tailed unpaired Student's t test in (B and F), two‐way analysis of variance (ANOVA) followed by Dunn‐Šídák post hoc analysis in (D), and one‐way ANOVA followed by Tukey's post hoc analysis in (H). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Expressing, Transgenic Assay, Ubiquitin Proteomics, Western Blot, Immunofluorescence, Staining, Marker, Comparison, Fluorescence, Mutagenesis, Control, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Astrocytic PYGM (glycogen phosphorylase, muscle associated) ablation exacerbates behavioral deficits in tau P301S transgenic (PS19) mice. (A) Experimental timeline for tamoxifen administration and behavioral tests. Four genotypes of mice were studied: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1L1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1L1 ‐cre ER ;PS19 (AcKO; PS19). (B‐D) Mice were subjected to the Morris Water Maze test to compare the escape latency to reach the platform during a 5‐day training phase (B), and the number of platform region crossings (C) and the average swimming speed (D) during the probe test. (E) Mice were studied for their spontaneous alternations in the Y‐maze test. (F, G) Mice were subjected to the fear conditioning test to compare the percentage of freezing time in the cued (F) and contextual (G) phases. (H) The body weights of mice were compared. (I, J) Representative images of the hindlimb clasping of different mice (I) and the comparison of hindlimb clasping scores (J). Data are presented as mean ± SEM. p values were determined by two‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (B), and one‐way ANOVA followed by Tukey's post hoc analysis in (C‐H, and J). n = 14 mice per group for all these tests. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Transgenic Assay, Comparison

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Astrocytic PYGM (glycogen phosphorylase, muscle associated) ablation exacerbates tauopathy‐related pathologies in tau P301S transgenic (PS19) mice. (A, B) Immunofluorescence staining of NeuN (red) (A) and quantitative analysis (B) in brain sections from different mice. The nuclei were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). Scale bars: 50 µm. n = 6 per group. (C, D) Immunofluorescence staining of phosphorylated tau at sites T231/S235 (red) (C) and quantitative analysis (D) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 8 per group. (E, F) Immunofluorescence staining of glial fibrillary acidic protein (GFAP; red) (E) and quantitative analysis (F) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 6 per group. (G, H) Immunofluorescence staining of Iba1 (red) (G) and quantitative analysis (H) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 6 per group. Data are presented as the mean ± SEM. p values were determined by one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (B, F, and H), and two‐tailed unpaired Student's t test in (D). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Transgenic Assay, Immunofluorescence, Staining, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Astrocytic PYGM (glycogen phosphorylase, muscle associated) overexpression attenuates cognitive and motor deficits in tau P301S transgenic (PS19) mice. (A) Experimental timeline for AAV delivery and behavioral tests. (B‐D) Mice were subjected to the Morris Water Maze test to compare the escape latency during a 5‐day training phase (B), and the number of platform region crossings (C) and the average swimming speed (D) during the probe test. (E) Mice were studied for their spontaneous alternations in the Y‐maze test. (F, G) Mice were subjected to the fear conditioning test to compare the percentage of freezing time in the cued (F) and contextual (G) phases. (H) The body weights of mice were compared. (I, J) Representative images of the hindlimb clasping (I) and the comparison of hindlimb clasping scores (J). Data are presented as mean ± SEM. p values were determined by one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (C‐H, and J), and two‐way ANOVA followed by Tukey's post hoc analysis in (B). n = 9 mice per group for all these tests. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Over Expression, Transgenic Assay, Comparison

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Astrocytic PYGM (glycogen phosphorylase, muscle associated) overexpression alleviates tauopathy‐related pathologies in tau P301S transgenic (PS19) mice. (A, B) Immunofluorescence staining of NeuN (green) (A) and quantitative analysis (B) in brain sections from different mice. The nuclei were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). Scale bars: 50 µm. n = 8 per group. (C, D) Immunofluorescence staining of phosphorylated tau at sites T231/S235 (green) (C) and quantitative analysis (D) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 8 per group. (E, F) Immunofluorescence staining of glial fibrillary acidic protein (GFAP; green) (E) and quantitative analysis (F) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 6 per group. (G, H) Immunofluorescence staining of Iba 1 (green) (G) and quantitative analysis (H) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 6 per group. Data are presented as the mean ± SEM. p values were determined by one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (B, F, and H), and two‐tailed unpaired Student's t test in (D). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Over Expression, Transgenic Assay, Immunofluorescence, Staining, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: PYGM (glycogen phosphorylase, muscle associated) deficiency disrupts glycogen‐energy metabolism in astrocytes. (A) Cultured primary astrocytes from wild‐type (WT) mice were infected with lentiviruses expressing shRNA targeting Pygm (shPygm) or control shRNA (shNC). The mRNA levels of Pygm , Pygb , Pygl , Ppp1r3c , Gys1 , and Gyg in shNC and shPygm astrocytes were analyzed by quantitative real‐time polymerase chain reaction (qRT‐PCR) for comparison. n = 6 experiments. (B, C) Periodic acid Schiff (PAS) staining of glycogen (magenta) in shNC and shPygm astrocytes (B) and quantitative analysis (C). Scale bars: 100 µm. n = 5 experiments. (D) Glycogen concentrations in shNC and shPygm astrocytes were assayed by a glycogen assay kit for comparison. n = 6 experiments. (E, F) Glycolysis capacity assessed by real‐time extracellular acidification rate (ECAR) monitoring in shNC and shPygm astrocytes (E) and corresponding quantitative comparisons (F). n = 12 experiments. (G) Lactate levels in the cell lysates and extracellular medium from shNC and shPygm astrocytes were quantified by a colorimetric assay kit for comparison. n = 4 experiments. (H) Comparison of lactate concentrations in the cerebrospinal fluid (CSF) of WT mice with astrocytic PYGM overexpression (WT‐OE) and their controls (WT‐NC). n = 7 per group. (I) Comparison of lactate concentrations in the CSF of PS19 mice with (OE) or without (NC) PYGM overexpression. n = 6 mice per group. (J, K) Mitochondrial respiration assessed by real‐time oxygen consumption rate (OCR) monitoring in shNC and shPygm astrocytes (J) and corresponding quantitative comparisons (K). n = 12 experiments. (L, M) shNC and shPygm astrocytes were stained with DCFH‐DA and analyzed by flow cytometry (L). shNC astrocytes without DCFH‐DA staining were used as a negative control. ROS levels were quantified and compared based on the mean fluorescence intensity of DCFH‐DA‐positive cells (M). n = 5 experiments. Data are presented as mean ± SEM. p values were determined by two‐tailed unpaired Student's t test in (A, C, D, F‐H, K, M) and one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (I). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Cell Culture, Infection, Expressing, shRNA, Control, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Comparison, Staining, Colorimetric Assay, Over Expression, Flow Cytometry, Negative Control, Fluorescence, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Astrocytic PYGM (glycogen phosphorylase, muscle associated) deficiency causes neuronal energetic impairment and aggravated Tau phosphorylation. (A) The conditioned media treatment scheme: conditioned media (ACM) collected from shNC or shPygm astrocytes were centrifuged at 1000 g for 10 minutes. The supernatants were used to treat wild‐type (WT) or tau P301S transgenic (PS19) primary neurons for 3 days. (B, C) Glycolysis capacity measured by real‐time extracellular acidification rate (ECAR) monitoring in neurons treated with shNC or shPygm ACM (B) and corresponding quantitative comparisons (C). n = 16 experiments. (D, E) Mitochondrial respiration measured by real‐time oxygen consumption rate (OCR) monitoring in neurons treated with shNC or shPygm ACM (D) and corresponding quantitative comparisons (E). n = 15 experiments. (F, G) Immunoblot (F) and quantitative analysis (G) of total tau and phosphorylated tau at sites T212, T231/S235, and T205 in neurons treated with shNC or shPygm ACM. n = 6 experiments. Data are presented as mean ± SEM. p values were determined by two‐tailed unpaired Student's t test in (G), and one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (C and E). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Phospho-proteomics, Transgenic Assay, Western Blot, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Astrocytic PYGM attenuates tau pathology by promoting lactate‐mediated neuroprotection

doi: 10.1002/alz.71202

Figure Lengend Snippet: Lactate supplementation mitigates tau‐related pathologies and cognitive deficits in tau P301S transgenic (PS19) mice. (A‐C) Immunofluorescence staining of Tuj1 (white) in wild‐type (WT) and PS19 primary neurons treated with PBS or lactate for 3 days (A). Scale bars: 20 µm. Quantitative comparisons of dendritic complexity using the Sholl analysis (B) and total dendritic branch length (C). n = 6 experiments with 50 neurons studied per group. (D, E) Immunoblot (D) and quantitative analysis (E) of total tau and phosphorylated tau at sites T212, T231/S235, and T205 in WT and PS19 neurons treated with phosphate buffered saline (PBS) or lactate. n = 4 experiments. (F) Schematic timeline for the in vivo L‐lactate administration and subsequent behavioral tests. (G) Mice were studied for their spontaneous alternations in the Y‐maze test. n = 12 mice per group. (H, I) Mice were subjected to the fear conditioning test to compare the percentage of freezing time in the cued (H) and contextual (I) phases. n = 12 mice per group. (J, K) Immunofluorescence staining of NeuN (green) (J) and quantitative analysis (K) in brain sections from different mice. The nuclei were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). Scale bars: 50 µm. n = 7 per group. (L, M) Immunofluorescence staining of phosphorylated tau at sites T231/S235 (green) (M) and quantitative analysis (N) in brain sections from different mice. The nuclei were counterstained with DAPI (blue). Scale bars: 100 µm. n = 7 per group. Data are presented as mean ± SEM. p values were determined by one‐way analysis of variance (ANOVA) followed by Tukey's post hoc analysis in (C, G‐I, and K), two‐way ANOVA followed by Tukey's post hoc analysis in (B), and two‐tailed unpaired Student's t test in (E and M). ns, not significant; * p < 0.05; ** p < 0.01; **** p < 0.0001.

Article Snippet: The resultant Pygm flox/flox ; Aldh1l1 ‐cre ER mice were further crossed with PS19 mice (Jackson Laboratory, Stock No: 008169; expressing human mutant tau P301S ) to generate four experimental groups: Pygm flox/flox (Ctrl), Pygm flox/flox ; Aldh1l1 ‐cre ER (AcKO), Pygm flox/flox ;PS19 (PS19), and Pygm flox/flox ; Aldh1l1 ‐cre ER ;PS19 (AcKO;PS19).

Techniques: Transgenic Assay, Immunofluorescence, Staining, Western Blot, Saline, In Vivo, Two Tailed Test

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of PLIN2 + lipid droplets (green), NeuN + neurons (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4 and PS19/E3 mice. b, Quantification of percent PLIN2 + area coverage in NeuN + neurons. c, Quantification of percent PLIN2 + area coverage in Iba1 + microglia. d, Representative immunofluorescent images of BODIPY + neutral lipids (green), DAPI + nuclei (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4 and PS19/E3 mice. a and d, Scale bars, 40μm. e, Quantification of percent BODIPY + area coverage in the hippocampal DG. f-i, Correlations between BODIPY + % area in DG and Iba1 + % area ( f ), AT8 + % area ( g ), NeuN + % area ( h ), and hippocampal volume ( i ) in PS19/E4 mice. j, Representative immunofluorescent images of BODIPY-C11 + peroxidized lipids (green), BODIPY-C11 + non-peroxidized lipids (red), Iba1 + microglia (blue), and GFAP + astrocytes (white) in PS19/E4 mice. Scale bars, 20μm k, Quantifications of BODIPY-C11 + peroxidized lipids in Iba1 + cells and GFAP + cells in PS19/E4 mice. l, Quantifications of BODIPY-C11 + non-peroxidized lipids in Iba1 + cells and GFAP + cells in PS19/E4 mice. Quantified data in b,c,e,k,l are represented as mean ± s.e.m., unpaired two-sided t-test. For all experiments, PS19/E4, n = 35; and PS19/E3 n = 42. Pearson’s correlation analysis (two-sided) used for f-i .

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: Correlations between PLIN2 + lipid droplet (LD) % area in Iba1 + microglia and Iba1 + % area in the hippocampus ( n = 34) ( a ), AT8 + % area in the hippocampus ( n = 34) ( b ), NeuN + % area in the hippocampus ( n = 35) ( c ), and hippocampal volume ( n = 33) ( d ) in PS19/E4 mice ( n = 35). e-h, Correlations between PLIN2 + LD % area in NeuN + neurons and Iba1 + % area ( e ), AT8 + % area ( f ), NeuN + % area ( g ), and hippocampal volume ( h ) in PS19/E4 mice. i-l, Correlations between PLIN2 + LD % area in Iba1 + microglia and Iba1 + % area ( n = 39) ( i ), AT8 + % area ( n = 31) ( j ), NeuN + % area ( n = 39) ( k ), and hippocampal volume ( n = 39) ( l ) in PS19/E3 mice. m-p, Correlations between PLIN2 + LD % area in NeuN + neurons and Iba1 + % area ( m ), AT8 + % area ( n ), NeuN + % area ( o ), and hippocampal volume ( p ) in PS19/E3 mice ( n = 42). Pearson’s correlation analysis (two-sided) was used in a-p .

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of PLIN2 + LD (green), NeuN + neurons (white), and Iba1 + microglia (red) in PS19/E4-mice at 4.0, 6.2, 7.8, and 10.0 months of age. Scale bars, 40μm. b, Correlation between age (in months) and PLIN2 + LDs in NeuN + neurons. Pearson’s correlation analysis (two-sided). c, Correlation between age (in months) and PLIN2 + LDs in Iba1 + microglia. Nonlinear regression R 2 . All scale bars, 40μm. For b,c PS19/E4 mice, n = 16

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of BODIPY + neutral lipids and GFAP + astrocyte in the hippocampal DG of PS19/E4 and PS19/E3 mice at 10 months of age. b-e, Correlations between BODIPY + neutral lipids in DG (% area) and Iba1 + % area ( n = 42) ( b ), AT8 + % area ( n = 31) ( c ), NeuN + % area ( n = 39) ( d ), and hippocampal volume ( n = 39) ( e ) in PS19/E3 mice, assessed via Pearson’s correlation analysis (two-sided). f, Representative immunofluorescent images of BODIPY + neutral lipids (green), DAPI + nuclei (white), and Iba1 + microglia (red) in the hippocampal DG of PS19/E4 mice at 4.0, 6.2, 7.8, and 10.0 months of age. Scale bars, 25μm. g, Correlation between age (in months) and BODIPY + neutral lipids in the hippocampal DG of PS19/E4 mice ( n = 20). Nonlinear regression R 2 . h-j, Representative immunofluorescent images of Rab5 + early endosomes (red, h ), Rab7 + late endosomes (red, i ), and LAMP1 + lysosomes (red, j ) with Iba1 + microglia (blue) and BODIPY + neutral lipid (green) in PS19/E4 mice. All scale bars, 40μm. k, Quantification of BODIPY + marker inside LAMP1 + stain ( n = 15).

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques: Marker, Staining

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of PLIN2 + lipid droplets (green), NeuN + neurons (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4, PS19/E4/Syn1-Cre, and PS19/NSE-E4 mice. Scale bars, 40μm. b, Quantification of percent PLIN2 + area coverage in NeuN + neurons. c, Quantification of percent PLIN2 + area coverage in Iba1 + microglia. d, Representative immunofluorescent images of BODIPY + neutral lipids (green), DAPI + nuclei (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4, PS19/E4/Syn1-Cre, and PS19/NSE-E4 mice. Scale bars, 40μm. e, Quantification of percent BODIPY + area coverage in the hippocampal DG mask. Quantified data in b-e represented as mean ± s.e.m., unpaired two-sided t-test. f-i, Correlations between BODIPY + % area in DG and Iba1 + % area ( f ), AT8 + % area ( g ), NeuN + % area ( h ), and hippocampal volume ( i ) in PS19/NSE-E4 mice. j, Representative immunofluorescent images of BODIPY-C11 + peroxidized lipids (green), BODIPY-C11 + non-peroxidized lipids (red), Iba1 + microglia (blue), and GFAP + astrocytes (white) in PS19/E4 mice. Scale bars, 20μm. k, Quantifications of BODIPY-C11 + peroxidized lipids in Iba1 + microglia and GFAP + astrocytes in PS19/NSE-E4 mice. l, Quantifications of BODIPY-C11 + non-peroxidized lipids in Iba1 + microglia and GFAP + astrocytes in PS19/NSE-E4 mice. Quantified data in k and l are represented as mean ± s.e.m., one-way analysis of variance (ANOVA) with Tukey’s post hoc multiple comparisons test. For all experiments, PS19/E4, n = 35; PS19/E3 n = 42; PS19/E4/Syn1-Cre n = 29; and PS19/NSE-E4 n = 18. Pearson’s correlation analysis (two-sided) used for f-i .

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: Correlations between BODIPY + neutral lipids in DG (% area) and Iba1 + % area ( n = 29) ( a ), AT8 + % area ( n = 25) ( b ), NeuN + % area ( n = 29) ( c ), and hippocampal volume ( n = 24) ( d ) in PS19/E4/Syn1-Cre mice. e-h, Correlations between PLIN2 + lipid droplets (LDs) (% area) in Iba1 + microglia and Iba1 + % area ( n = 25) ( e ), AT8 + % area ( n = 25) ( f ), NeuN + % area ( n = 24) ( g ), and hippocampal volume ( n = 24) ( h ) in PS19/E4/Syn1-Cre mice. i-l, Correlations between PLIN2 + LDs (% area) in NeuN + neurons and Iba1 + % area ( n = 25) ( i ), AT8 + % area ( n = 25) ( j ), NeuN + % area ( n = 24) ( k ), and hippocampal volume ( n = 24) ( l ) in PS19/E4/Syn1-Cre mice. m-p, Correlations between PLIN2 + LDs (% area) in Iba1 + microglia and Iba1 + % area ( n = 29) ( m ), AT8 + % area ( n = 15) ( n ), NeuN + % area ( n = 18) ( o ), and hippocampal volume ( n = 17) ( p ) in PS19/NSE-E4 mice ( n = 18). q-t, Correlations between PLIN2 + LDs (% area) in NeuN + neurons and Iba1 + % area ( n = 18) ( q ), AT8 + % area ( n = 15) ( r ), NeuN + % area ( n = 18) ( s ), and hippocampal volume ( n = 17) ( t ) in PS19/NSE-E4 mice. Pearson’s correlation analysis (two-sided) was used for a-t .

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of Olig2 + (red) oligodendrocytes co-localizing with BODIPY + neutral lipids (green) in the hippocampal DG of 10-month-old PS19/E4, PS19/E4/Syn1-Cre, and PS19/NSE-E4 mice. Yellow arrows in zoomed in images represent co-localization of neutral lipids inside Olig2 + oligodendrocytes. Yellow arrows in zoomed-in images represent Olig2 + oligodendrocytes containing BODIPY + neutral lipids. Scale bars, 40μm. b, Quantification of percent BODIPY + area coverage in Olig2 + oligodendrocytes. (PS19/E4, n = 13; PS19/E3 n = 12; PS19/E4/Syn1-Cre n = 5, and PS19/NSE-E4 n = 7). c, Representative immunofluorescent images of BODIPY + neutral lipids (green), Olig2 + oligodendrocytes (red), and Iba1 + microglia (white) in the hippocampal DG of 10-month-old PS19/E4, PS19/E4/Syn1-Cre, and PS19/NSE-E4 mice. Yellow arrows in zoomed-in images represent Iba1 + microglia containing both Olig2 + oligodendrocytes and BODIPY + neutral lipids. Scale bars, 40μm. d, Quantification of percent of Iba1 + microglia that contain both Olig2 + oligodendrocytes and BODIPY + neutral lipids, expressed as a percentage of total Iba1 + microglia. (PS19/E4 n = 12; PS19/E3 n = 12; PS19/E4/Syn1-Cre n = 8, and PS19/NSE-E4 n = 8). Quantified data in b,d are represented as mean ± s.e.m., one-way analysis of variance (ANOVA) with Tukey’s post hoc multiple comparisons test.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of TUJ1 + neurons (green), GFAP + astrocytes (red), and Iba1 + microglia (white) in primary mouse neuron culture. Iba1 + (white channel) shows minimal signal due to background debris. b, Representative immunofluorescent images of P2RY12 + microglia (green), CD68 + microglia (red), and Iba1 + microglia (white) in primary mouse microglia culture. c, Representative immunofluorescent images of PLIN2 + lipid droplets (LDs, green) in primary PS19/E4 Iba1 + microglia (white) treated with neuron-conditioned media (either PS19/E4 neuron CM or PS19/E4/Syn1-Cre neuron CM) for 72 hours. d, Quantification of percent PLIN2 + area coverage in Iba1 + microglia (PS19/E4 neuron CM, n = 4; PS19/E4/Syn1-Cre neuron CM, n = 4). e, Representative immunofluorescent images of BODIPY + neutral lipids (green), PLIN2 + LDs (red) in TUJ1 + neurons (white) from PS19/E4 or PS19/E4/Syn1-Cre mice. f, Quantification of percent BODIPY + area coverage in TUJ1 + neurons (PS19/E4 neuron, n = 4; PS19/E4/Syn1-Cre neuron, n = 5). g, Quantification of percent PLIN2 + area coverage in TUJ1 + neurons (PS19/E4 neuron, n = 4; PS19/E4/Syn1-Cre neuron, n = 5). h, Representative brightfield image of morphological observation of mature oligodendrocytes. Scale bar, 40μm. i, Representative immunofluorescent images of MBP + (red) and Olig2 + (white) oligodendrocytes in PS19/E4. All scale bars, 10μm unless otherwise stated. Quantified data are represented in d,f,g as mean ± s.e.m., unpaired two-sided t-test.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques:

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of BODIPY + neutral lipids (green) and Iba1 + microglia (white) from PS19/E4 mice, treated with conditioned media (CM) from neurons of PS19/E4 or PS19/E4/Syn1-Cre mice for 72 hours. b, Quantification of percent BODIPY + area coverage in Iba1 + microglia (MG) from PS19/E4 mice (PS19/E4 neuron CM, n = 8; PS19/E4/Syn1-Cre neuron CM n = 4). c, Representative immunofluorescent images of BODIPY + neutral lipids (green), Iba1 + microglia (white), and TUJ1 + neurons (red) of PS19/E4 microglia co-cultured with PS19/E4 or PS19/E4/Syn1-Cre neurons for 72 hours. Arrows point to BODIPY in Iba1 + microglia. d, Quantification of percent BODIPY + area coverage in Iba1 + microglia (MG) from PS19/E4 mice (PS19/E4 neuron, n = 4; PS19/E4/Syn1-Cre neuron, n = 4). e, Representative immunofluorescent images of BODIPY-C16 + lipids (green), Iba1 + microglia (white), and TUJ1 + neurons (red) from the lipid transfer assay with PS19/E4 microglia and PS19/E4 neurons. Yellow arrows point towards BODIPY-C16-lipids transferred from neurons to microglia and white arrow points toward BODIPY-C16-lipids remaining in neurons. ( n = 4). f, Quantification of BODIPY-C16-lipids in TUJ1 + neurons (checkered) and Iba1 + microglia (stripe). (PS19/E4 neuron, n = 4; PS19/E4/Syn1-Cre neuron, n = 4). g, Quantification of count represented in percent of Iba1 + microglia containing TUJ1 + (red dots), BODIPY + (green dots), and TUJ1 + and BODIPY + (yellow dots) signals. (PS19/E4, n = 4). h, Representative immunofluorescent images of peroxidized BODIPY-C11-lipids (B-C11, green), non-peroxidized BODIPY-C11-lipids (B-C11, red), Iba1 + microglia (white), and TUJ1 + neurons (blue) in mixed neuron-microglia co-culture. i, Quantifications of BODIPY-C11 + peroxidized lipids in Iba1 + microglia. (PS19/E4 neuron, n = 5; PS19/E4/Syn1-Cre neuron, n = 4). (PS19/E4 neuron, n = 5; PS19/E4/Syn1-Cre neuron, n = 4). j, Representative immunofluorescent images of BODIPY + neutral lipids (green), MBP+ oligodendrocytes (red), Iba1 + microglia (white) and TUJ1 + neurons (blue) in mixed culture of PS19/E4 microglia, PS19/E4 oligodendrocytes, and either PS19/E4 or PS19/E4/Syn1-Cre neurons. Scale bars, 10μm. k , Quantification of percent BODIPY + area coverage in Iba1 + microglia in the mixed culture of PS19/E4 microglia, PS19/E4 oligodendrocytes, and either PS19/E4 or PS19/E4/Syn1-Cre neurons. (PS19/E4 neuron, n = 10; PS19/E4/Syn1-Cre neuron, n = 6). l, Quantification of count represented in percent of Iba1 + microglia containing both MBP + oligodendrocytes and BODIPY + neutral lipids, expressed as a percentage of total Iba1 + microglia. (PS19/E4-Neuron Condition, n = 10; PS19/E4/Syn1-Cre Neuron Condition n = 5). Quantified data in b,d,f,g,i,k,l represented as mean ± s.e.m., unpaired two-sided t-test. In a,c,e,h,j all scale bars, 10μm.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques: Cell Culture, Co-Culture Assay

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Representative immunofluorescent images of Iba1 + microglia (red) in 10-month-old PS19/E4 mice treated with either control or PLX5622 (PLX) chow for 3 months. b, Quantification of percent Iba1 + area coverage in PS19/E4-Control or PS19/E4-PLX mice. c, Representative immunofluorescent images of GFAP + astrocytes (white) in 10-month-old PS19/E4 mice treated with either control or PLX5622 chow for 3 months. d, Quantification of percent GFAP + area coverage in PS19/E4-Control or PS19/E4-PLX mice. e, Representative immunofluorescent images of PLIN2 + lipid droplets (green), NeuN + neurons (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4-Control or PS19/E4-PLX mice. f, Quantification of percent PLIN2 + area coverage in Iba1 + microglia. g, Representative immunofluorescent images of PLIN2 + lipid droplets (green), NeuN + neurons (white), and Iba1 + microglia (red) in the hippocampal DG of 10-month-old PS19/E4-Control or PS19/E4-PLX mice. h, Quantification of percent BODIPY + area coverage in the hippocampal DG of PS19/E4-Control or PS19/E4-PLX mice. In a,c,e,g, all scale bars, 40μm. In f,h, each color corresponds to one mouse, with two brain sections per mouse for PS19/E4-Control mice ( n = 4) and PS19/E4-PLX mice ( n= 6). Quantified data in b,d,f,h are represented as mean ± s.e.m., unpaired two-sided t-test.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques: Control

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, UMAP plot of all 36 distinct cell clusters in the hippocampi of 10-month-old PS19/E4, PS19/E3, PS19/E4/Syn1-Cre, and PS19/NSE-E4 mice ( n = 4 for each genotype). b, Genotype-split UMAP plot highlighting cells in excitatory neuron clusters 4, 11, and 17 for each genotype group. c, Box plots of proportion of cells from each sample in excitatory neuron clusters 4, 11, and 17. The lower, middle, and upper hinges of the box plot correspond to the 25 th , 50 th , and 75 th percentiles, respectively. The upper whisker of the box plot extends from the upper hinge to the largest value no further than 1.5 x IQR from the upper hinge. The lower whisker extends from the lower hinge to the smallest values at most 1.5 x IQR from the lower hinge. IQR, interquartile range or distance between 25 th and 75 th percentiles. Data beyond the end of the whiskers are the outlier points. The LORs are the mean ± s.e.m. estimates of LOR for these clusters, which represents the change in the log odds of cells per sample from PS19/E3, PS19/E4/Syn1-Cre, or PS19/NSE-E4 mice belonging to the respective clusters compared to the log odds of cells per sample from PS19-E4 mice. PS19/E3 is represented in yellow, PS19/E4 is represented in red, PS19/E4/Syn1-Cre is represented in green, and PS19/NSE-E4 is represented in pink. LOR significant differences from either PS19/E4 or PS19/E3 are highlighted in corresponding color of the genotype (e.g. if PS19/NSE-E4 is significantly different, text is colored pink). d, Heat map plot of log odds ratio (LOR) per unit change in each pathological measurement for clusters 4, 11, and 17. The LOR represents the mean estimate of the change in the log odds of cells per sample from a given animal model, corresponding to a unit change in a given histopathological parameter. Associations with pathologies are colored (negative associations, blue; positive associations, red). P values in c are from fits to a GLMM_AM, and p values in d are from fits to a GLMM_histopathology; the associated tests are two-sided. All error bars represent s.e.m. * p < 0.05, ** p <0.01, *** p <0.001. e,g,i, Volcano plot for the top 40 (20 up and 20 down) differentially expressed genes (DEGs) of excitatory neuron cluster 4 ( e ), 11 ( g ), and 17 ( i ) for PS19/E4 versus PS19/E4/Syn1-Cre. f,h,j, Volcano plot for the top 20 DEGs of excitatory neuron cluster 4 ( f ), 11 ( h ), and 17 ( j ) for PS19/NSE-E4 versus PS19/E4/Syn1-Cre.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques: Whisker Assay, Animal Model, Histopathology

Journal: bioRxiv

Article Title: Neuronal APOE4 drives damaging lipid accumulation via contact-dependent neuron-oligodendrocyte-microglia interaction in Alzheimer’s disease

doi: 10.64898/2025.12.04.692390

Figure Lengend Snippet: a, Genotype-split UMAP plot highlighting cells in oligodendrocyte cluster 7 and microglial clusters 10 and 14 for each genotype group. b, Box plot of proportion of cells from each sample in cluster 7. c, Heat map plot of log odds ratio (LOR) per unit change in each pathological measurement for clusters 7, 10, and 14. The LOR represents the mean estimate of the change in the log odds of cells per sample from a given animal model, corresponding to a unit change in a given histopathological parameter. Associations with pathologies are colored (negative associations, blue; positive associations, red). d,e, Volcano plot for the top 40 (20 up and 20 down) differentially expressed genes (DEGs) of oligodendrocyte cluster 7 for PS19/E4 versus PS19/E4/Syn1-Cre ( d ) and for PS19/NSE-E4 vs PS19/E4/Syn1-Cre ( e ). f, Box plot of proportion of cells from each sample in cluster 10. g,h, Volcano plot for the top 40 (20 up and 20 down) DEGs of microglia cluster 10 for PS19/E4 versus PS19/E4/Syn1-Cre ( g ) and for PS19/NSE-E4 vs PS19/E4/Syn1-Cre ( h ). i, Box plot of proportion of cells from each sample in cluster 14. j,k, Volcano plot for the top 40 (20 up and 20 down) DEGs of microglia cluster 14 ( j ) for PS19/E4 versus PS19/E4/Syn1-Cre and for PS19/NSE-E4 vs PS19/E4/Syn1-Cre ( k ). For b,f,i, the lower, middle, and upper hinges of the box plot correspond to the 25 th , 50 th , and 75 th percentiles, respectively. The upper whisker of the box plot extends from the upper hinge to the largest value no further than 1.5 x IQR from the upper hinge. The lower whisker extends from the lower hinge to the smallest values at most 1.5 x IQR from the lower hinge. IQR, interquartile range or distance between 25 th and 75 th percentiles. Data beyond the end of the whiskers are the outlier points. The LORs are the mean ± s.e.m. estimates of LOR for these clusters, which represents the change in the log odds of cells per sample from PS19/E3, PS19/E4/Syn1-Cre, or PS19/NSE-E4 mice belonging to the respective clusters compared to the log odds of cells per sample from PS19-E4 mice. PS19/E3 is represented in yellow, PS19/E4 is represented in red, PS19/E4/Syn1-Cre is represented in green, and PS19/NSE-E4 is represented in pink. P values in b,f,i are from fits to a GLMM_AM, and p values in c are from fits to a GLMM_histopathology; the associated tests are two-sided. All error bars represent s.e.m. * p < 0.05, ** p <0.01, *** p <0.001.

Article Snippet: The resulting E/Cre mice were subsequently bred with tau-P301S (PS19) transgenic mice (B6;C3-Tg(Prnp-MAPT*P301S)PS19Vle/J; The Jackson Laboratory, 008169), which express the human 1N4R tau isoform with the P301S mutation under the PrP promoter, to generate PS19/E3 and PS19/E4 mice either with or without Syn1-Cre, as previously described .

Techniques: Animal Model, Whisker Assay, Histopathology

Journal: Acta Neuropathologica Communications

Article Title: Telomere-driven senescence accelerates tau pathology, neuroinflammation and neurodegeneration in a tauopathy mouse model

doi: 10.1186/s40478-025-02118-5

Figure Lengend Snippet: Telomere shortening in the brains of G3Terc −/− and G2Terc −/− mice triggers the activation of classical senescence markers. ( a ) Relative telomere length detected by qPCR analysis using hippocampal DNA extracts from WT and successive generations (G1-G3) of Terc −/− mice at 5 months of age. The average telomere length was calculated as the ratio (T/S) of telomere repeat copy number (T) to a single-copy gene (S = 36B4). **P < 0.01, ***P < 0.001 (One-way ANOVA with Tukey’s post-hoc analysis, n = 4–7 mice/group). ( b-c ) Relative levels of the DNA damage biomarker γH2AX detected by Western Blot analysis using hippocampal protein extracts from 5-6-month-old WT and G3Terc −/− mice ( b ) and 9-month-old WT and G2Terc −/− mice ( c ). **P < 0.01, ***P < 0.001 (two-tailed Student’s t -test, n = 11–13). ( d-e ) mRNA levels of the SASP factors Il1b and Cxcl1, and the cyclin-dependent kinase inhibitors p19 Arf (p19) and p21 Waf1/cip1 (p21), were measured by RT-qPCR in hippocampal extracts from 5-month-old WT and G3Terc −/− mice ( d ) and 9-month-old WT and G2Terc −/− mice ( e ). *P < 0.05, **P < 0.01, ***P < 0.001 (two-tailed Student’s t- test or Mann-Whitney’s test, n = 5–8 mice/group)

Article Snippet: Heterozygous Terc knockout mice (referred to as Terc +/− , Strain #004132, The Jackson Laboratory), carrying a germline deletion of the telomerase RNA component Terc [ ], and hemizygous Tau P301S (PS19 line) transgenic mice (referred to as Tau Tg , Strain #008169, The Jackson Laboratory), overexpressing the 1N4R human tau isoform with the P301S mutation found in frontotemporal dementia with parkinsonism-17 (FTDP-17) [ ], were purchased and maintained on a C57BL/6 genetic background for a minimum of 6 generations.

Techniques: Activation Assay, Biomarker Discovery, Western Blot, Two Tailed Test, Quantitative RT-PCR